Effects of Three Consecutive Days of Morphine or Methadone Administration on Analgesia and Open-Field Activity in Mice with Ehrlich Carcinoma.

This study assessed the exploratory behavioral responses in BALB/c mice inoculated with Ehrlich ascitic carcinoma after 3 consecutive days of treatment with morphine or methadone. Fifty-three female mice, 60 ± 10 d old, were used. Seven days after intraperitoneal tumor inoculation (2 × 106 cells), the animals were randomized into 7 groups: morphine 5 mg/kg (MO5), morphine 7.5 mg/kg (MO7.5), morphine 10 mg/kg (MO10), methadone 2.85 mg/kg (ME2.85), methadone 4.3 mg/kg (ME4.3), methadone 5.7 mg/kg (ME5.7), and 0.9% NaCl (Saline) (n = 7). Drug treatments were administered subcutaneously every 6 h for 3 d. The animals were evaluated for analgesia using the mouse grimace scale (MGS) and for general activity using the open field test. The MGS was performed before tumor inoculation (day 0), on day 7 at 40, 90, 150, 240, and 360 min after drug injection, and on days 8 and 9 at 40, 150, 240, and 360 min after drug injection. The open field test was performed before tumor inoculation (day 0), on day 7 after inoculation at 40, 90, 150, 240, and 360 min after drug injection, and on days 8 and 9 after inoculation at 40, 150, and 360 min after drug injection. MGS results indicated that administration of morphine promoted analgesia for up to 240 min. Conversely, methadone reduced MGS scores only at 40 min. All tested doses promoted a significant dose-dependent increase in the total distance traveled and the average speed, and increase that was markedly pronounced on days 8 and 9 as compared with day 7. The frequencies of rearing and self-grooming decreased significantly after morphine or methadone administration. Despite the difference in analgesia, both drugs increased locomotion and reduced the frequency of rearing and self-grooming as compared with the untreated control animals.

Stereological analysis of the New Zealand rabbits (Oryctolagus cuniculus) placenta.

The onset of gestation is characterized by growth, morphological and functional changes of the placenta. We aim to evaluate the placental compartments in New Zealand rabbits by means of stereological methods. The fetal and maternal portion of placenta (12, 14, 18 and 20 gestational days) was randomly sampled for the stereological analysis. Histological sections were scanned to estimate fetal (labyrinth and junctional) and maternal (decidua) compartment volumes. The total volume of the placenta for the ages of 12, 14, 18 and 20 days was, respectively, 320 mm3, 340 mm3, 940 mm3 and 1300 mm3. The volume of the labyrinth was 56 mm3, 119 mm3, 231 mm3 and 481 mm3, respectively. The volume of junctional zone was 75 mm3, 76 cm3, 238 mm3 and 314 mm3, respectively. The volume of decidua was 174 mm3, 143 mm3, 469 mm3 and 504 mm3, respectively. We concluded that the rabbit´s placenta compartments varied according to the gestational period, increasing continuously over the 20 gestational days. However, on the onset of the development of the placenta the decidua presented faster growth, whereas after the 20 days of development, the labyrinth developed more quickly. This study represents an aid to the understanding of placentation in humans.

Characterization and Immunomodulation of Canine Amniotic Membrane Stem Cells.

PURPOSE: Amniotic membrane stem cells have a high capacity of proliferation, cell expansion, and plasticity, as well as immunomodulatory properties that contribute to maternal-fetal tolerance. Owing to the lack of research on human amniotic membrane at different gestational stages, the canine model is considered ideal because of its genetic and physiological similarities. We aimed to characterize the canine amniotic membrane (CAM) cell lineage in different gestational stages and evaluate the expression of immunomodulatory genes. MATERIALS AND METHODS: Twenty CAMs from early (20-30 days) (n=7), mid- (31-45 days) (n=7), and late gestation (46-63 days) (n=6) stages were studied. The cell features were assessed by cell viability tests, growth curve, colony-forming units, in vitro differentiation, cell labeling for different immunophenotypes, and pluripotent potential markers. The cells were subjected to RT-PCR and qPCR analysis to determine the expression of IDO, HGF, EGF, PGE2, and IL-10 genes. RESULTS: CAM cells exhibited a fibroblastoid morphology and adherence to plastic with an average cell viability of 78.5%. The growth curve indicated a growth peak in the second passage and we obtained an average of 138.2 colonies. Osteogenic, chondrogenic, and adipogenic lineages were confirmed by in vitro differentiation assays. Cellular immunophenotyping experiments confirmed the presence of positive mesenchymal markers (CD90 and CD105) and the low or negative expression of hematopoietic markers (CD45 and CD34). Qualitative analysis of the immunomodulatory functions indicated the expression of the IDO, HGF, EGF5, and PGE2 genes. When stimulated by interferon-gamma, CAM cells exhibited higher IDO levels throughout gestation. CONCLUSION: The CAMs from different gestational stages presented features consistent with mesenchymal stem cell lineage; better results were observed during the late gestation stage. Therefore, the gestational stage is a key factor that may influence the functionality of therapies when using fetal membrane tissues from different periods of pregnancy.

The timeline development of female canine germ cells.

During the sex differentiation, the primordial germ cells (PGCs) pass through a differentiation, becoming spermatogonial cells in males and oocytes in females. In this phase, there is difference in gene expression and differentiation potency between males and females. Specific cell markers have been essential in the PGC meiosis beginning and become oocyte cells. However, there are few studies about germline in domestic animals. The domestic dog (Canis lupus familiaris) is an interesting animal model to be used in the investigation about the mammal development because it has several biochemical and physiological similarities to humans. In addition, some additional investigations about dogs may contribute to a better understanding of the biology and genetic components, improving clinical veterinary and zoological sciences. Here, we elucidated by immunofluorescence and quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR), the dynamics of the expression of pluripotent (POU5F1 and NANOG) and germline (DDX4, DAZL and DPPA3) markers that are very important in the development of female canine germ cells during 35-50 days post-fertilization (dpf). The female canine germ cells were positive for pluripotent markers during middle developmental period. The number of DDX4, DAZL and DPPA3 cells increased along the germ cell maturation from 45 to 50 dpf. We provided an expression analysis of the pluripotent and germline markers in paraffin sections using the middle and later periods in female canine germ cells. The results can contribute the understanding about the timeline of each marker along the maturation of female canine germ cells. These results have a great significance to demonstrate the germ cell profile changes because it may allow the development of protocols about in vitro germ cell derivation.

Dynamics of male canine germ cell development.

Primordial germ cells (PGCs) are precursors of gametes that can generate new individuals throughout life in both males and females. Additionally, PGCs have been shown to differentiate into embryonic germ cells (EGCs) after in vitro culture. Most studies investigating germinative cells have been performed in rodents and humans but not dogs (Canis lupus familiaris). Here, we elucidated the dynamics of the expression of pluripotent (POU5F1 and NANOG), germline (DDX4, DAZL and DPPA3), and epigenetic (5mC, 5hmC, H3K27me3 and H3K9me2) markers that are important for the development of male canine germ cells during the early (22-30 days post-fertilization (dpf)), middle (35-40 dpf) and late (45-50 dpf) gestational periods. We performed sex genotype characterization, immunofluorescence, immunohistochemistry, and quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) analyses. Furthermore, in a preliminary study, we evaluated the capacity of canine embryo PGCs (30 dpf) to differentiate into EGCs. To confirm the canine EGCs phenotype, we performed alkaline phosphatase detection, immunohistochemistry, electron and transmission scanning microscopy and RT-qPCR analyses. The PGCs were positive for POU5F1 and H3K27me3 during all assessed developmental periods, including all periods between the gonadal tissue stage and foetal testes development. The number of NANOG, DDX4, DAZL, DPPA3 and 5mC-positive cells increased along with the developing cords from 35-50 dpf. Moreover, our results demonstrate the feasibility of inducing canine PGCs into putative EGCs that present pluripotent markers, such as POU5F1 and the NANOG gene, and exhibit reduced expression of germinative genes and increased expression of H3K27me3. This study provides new insight into male germ cell development mechanisms in dogs.

In vitro identification of a stem cell population from canine hair follicle bulge region.

Skin is an extensive and easily accessible organ possessing various cell types that are constantly renewed. Previous studies have suggested the presence of a stem cell niche at the bulge region of the hair follicle, which contains cells positive for CD200 and CD34. Thus, this study sought to identify these cell populations in canine skin cells using the following methods 1- collecting samples of adult and fetal skin and isolating and culturing these cells using a method of simple enzymatic digestion and 2- testing the cell cultures for CD200 and CD34 in vitro and comparing them with skin tissue samples (in situ). Immunofluorescence results were negative for both CD200 and CD34 in frozen and paraffin embedded tissue, whereas the analysis showed that cultured cells positive for CD34, CD200 and double positive cells could be visualized in different percentages. Additionally, the pluripotency marker OCT4 was positive in the isolated cells. Analysis of CD34, CD200 and OCT4 by RT-qPCR showed that there is expression in fetal and adult cells, although no difference was observed between groups. Our results suggest that bulge stem cells from both fetuses and adult dogs were reported with the use of CD34 and CD200 markers in this study, and further techniques for cell isolation and in vitro cultivation are needed in order to obtain enriched populations of skin stem cells in dogs.

Bone marrow stem cell applied in the canine veterinary clinics / Células tronco de medula óssea utilizadas na clínica veterinária canina

Cell therapy represents an old therapeutic practice initiated with the transfusion of whole blood in different clinical situations. There is now a breakthrough in the study of multipotent stem cell therapy because of its functionality in regeneration of tissues, which promotes attention of the scientific community. Bone marrow is one of the main sources of multipotent stem cells, composed by hematopoietic stem cells responsible for the renewal of the cellular components of the blood, and mesenchymal stem cells that aid in the regeneration of tissues. These cells have a strong potential for the treatment of several diseases, due their main characteristics such as high plasticity, capacity for self-renewal and immunomodulation. Although, there are many studies that show good results with the use of cell therapy as a form of treatment for several diseases, some studies still show inconclusive or unsatisfactory results. Therefore, the objective of this study was to review the application of bone marrow stem cells in the canine model since improvements on the knowledge of the technique are necessary to enable its applicability with safety and efficacy.(AU)

A terapia celular representa uma antiga prática terapêutica iniciada com a transfusão de sangue total em diferentes situações clínicas. Atualmente há um avanço no estudo da terapia com células-tronco mesenquimais por conta de sua funcionalidade na regeneração de tecidos, o que promove uma crescente atenção do meio científico. A medula óssea é uma das principais fontes de células-tronco multipotentes, no qual se encontram as células-tronco hematopoiéticas, responsável pela renovação dos componentes celulares do sangue, e as células-tronco mesenquimais que auxiliam na regeneração de tecidos. Essas células têm um forte potencial para o tratamento de diversas enfermidades, uma vez que possuem como principais características alta plasticidade, capacidade de auto renovação e imunomodulação. Apesar de haver muitos trabalhos que apresentam bons resultados com a utilização da terapia celular como forma de tratamento para diversas enfermidades, alguns estudos ainda demonstram resultados inconclusivos ou não satisfatórios, por isso, objetivou-se com este trabalho revisar a aplicação das células-tronco derivadas da medula óssea no modelo canino uma vez que é necessário melhorias sobre o conhecimento da técnica para que possibilite a sua aplicabilidade com segurança e eficácia.(AU)

Bioimpressão e produção de mini-órgãos com células tronco / Bioprinting with stem cells and production of mini-organs

A bioimpressão é considerada uma fonte promissora no desenvolvimento celular, e na produção de mini-órgãos, válulas, cartilagens que futuramente poderão ser utilizados na terapia para transplantes em animais e humanos. Assim, essa técnica poderá ser utilizada como uma terapia eletiva, no tratamento de injúrias e principalmente no tratamento de doenças crônico-degenerativas. Em humanos essa terapia está sendo pesquisada a fim de auxiliar a medicina no tratamento e regeneração de tecidos impressos a partir de arcabouços de células desenvolvidas a partir de células-tronco, biomateriais e impressões em 3D. O uso dessa tecnologia é também um auxiliar nas pesquisas oncológicas com o intuito de projetar e avaliar a proliferação celular de tumores, bem como a ação de novos medicamentos quimioterápicos. No entanto, a maior limitação para o uso da terapia utilizando-se a impressora de células, órgãos e tecidos em 3D é a falta de protocolos unificados com metodologias reprodutíveis e detalhadas; com o objetivo de viabilizar a utilização da impressora e a impressão de células, órgãos e tecidos em 3D. Dessa forma, esta revisão busca reunir as publicações mais atuais na área, as quais destacam os avanços no uso de bioimpressão com células-tronco, a fim de descrever as principais técnicas e os potenciais de utilização como alternativa terapêutica na medicina humana e veterinária.(AU)

The bioprinting is considered a promising source in cell development, and production of mini-organs, valves, cartilage that may eventually be used in therapy for transplantation in animals and humans. It can also be used as an elective therapy in the treatment of injuries and treatment of chronic degenerative diseases. In humans, this therapy is been studied mainly in the treatment and regeneration of tissues printed from scaffold cells developed from stem cells, biomaterials and impressions in 3D. This technology is also an aid for the study of the formation of tumors, in order to design and evaluate the cellular proliferation of the tumors and the action of new chemotherapy drugs. However, the main drawback to this therapy is the lack of standardized protocols with reproducible and detailed methodologies with the aim of enabling the use of bioprinting and printing cells, tissues and organs in 3D. Thus, this review seeks to bring together the most current publications of the bioprinting area in order to describe the technique and its potential use as a therapeutic alternative.(AU)

Maintenance of brazilian biodiversity by germplasm bank / Manutenção da biodiversidade brasileira através de bancos de germoplasma

Currently the importance of using alternative strategies for biodiversity conservation is emphasized and since the establishment of germplasm bank is an alternative to the conservation of endangered species. This is a technique of great importance for the maintenance of Brazilian fauna. Since the early70'sthere was a growing concern about the need to preserve essential genetic resources for food and agriculture, mainly for conservation of genetic material from farm animals. Thus was created the Brasilia Zoo, in July 2010, the first Germplasm Bank of Wild Animals in Latin America, as an alternative strategy for the conservation of threatened or endangered species, using both gametes and somatic cells and stem cells. Then we argue to create new banks or research networks among different regions with aimed to tissue preservation.

Atualmente, a importância do uso de estratégias alternativas para a preservação da biodiversidade é ressaltada e, visto que a criação de bancos de germoplasma é uma alternativa para a conservação de espécies ameaçadas, esta é uma técnica de suma importância para a manutenção da fauna brasileira. Desde o começo da década de 70 houve uma crescente preocupação sobre a necessidade de se preservar recursos genéticos essenciais para alimentação e agricultura, voltados principalmente, para a conservação de material genético de animais de produção. Deste modo, foi criado pelo Jardim Zoológico de Brasília, em julho de 2010, o primeiro Banco de Germoplasma de Animais Selvagens da América Latina, como uma estratégia alternativa para a conservação de espécies ameaçadas ou em perigo de extinção, utilizando tanto gametas como células somáticas e células-tronco. Com isto ponderamos na criação de novos bancos ou redes de pesquisa inter-regionais que foquem nesta preservação tecidual.

Profiles of Steroid Hormones in Canine X-Linked Muscular Dystrophy via Stable Isotope Dilution LC-MS/MS.

Golden retriever muscular dystrophy (GRMD) provides the best animal model for characterizing the disease progress of the human disorder, Duchenne muscular dystrophy (DMD). The purpose of this study was to determine steroid hormone concentration profiles in healthy golden retriever dogs (control group - CtGR) versus GRMD-gene carrier (CaGR) and affected female dogs (AfCR). Therefore, a sensitive and specific analytical method was developed and validated to determine the estradiol, progesterone, cortisol, and testosterone levels in the canine serum by isotope dilution liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). To more accurately understand the dynamic nature of the serum steroid profile, the fluctuating levels of these four steroid hormones over the estrous cycle were compared across the three experimental groups using a multivariate statistical analysis. The concentration profiles of estradiol, cortisol, progesterone, and testosterone revealed a characteristic pattern for each studied group at each specific estrous phase. Additionally, several important changes in the serum concentrations of cortisol and estradiol in the CaGR and AfCR groups seem to be correlated with the status and progression of the muscular dystrophy. A comprehensive and quantitative monitoring of steroid profiles throughout the estrous cycle of normal and GRMD dogs were achieved. Significant differences in these profiles were observed between GRMD and healthy animals, most notably for estradiol. These findings contribute to a better understanding of both dog reproduction and the muscular dystrophy pathology. Our data open new venues for hormonal behavior studies in dystrophinopathies and that may affect the quality of life of DMD patients.

Morfologia das glândulas salivares de glironia venusta Thomas, 1912 (didelphimorphia) / Morphology of salivary glands of the glironia venusta Thomas, 1912 (didelphimorphia)

O objetivo deste trabalho foi descrever a morfologia das glândulas salivares de Glironia venusta. Foi utilizado um exemplar coletado no resgate de fauna da Usina Hidroeletrica Teles Pires. Este foi fixado com solução de formaldeído a 10%. Para a análise macroscópica, as glândulas foram dissecadas e fotografadas in situ e para a microscopia foram retirados fragmentos das glândulas. Estes foram desidratados em concentrações crescentes de álcool, diafanizados em xilol, inclusos em parafina e corados com HE. G. venusta apresentou as glândulas mandibulares, parótidas, sublinguais, labiais e zigomáticas. As mandibulares se mostraram como estruturas alongadas, constituídas por lobos não septados e localizadas na porção anterior da região cervical. De formato triangular, a parótida estava localizada na depressão do masseter, ventral à cartilagem auricular. As sublinguais encontravam-se ventrais ao ramo da mandíbula, caudais ao digástrico. Foi observado na porção dorsoposterior à comissura labial, dorsal ao músculo orbicular da boca, as glândulas labiais, que apresentaram formato de "U". Já as glândulas zigomáticas acompanhavam a forma da porção anterior do arco zigomático, na margem infraorbital. As glândulas mandibulares e sublinguais apresentaram ácinos do tipo mucosos, com alguns ácinos serosos. A parótida era constituída por ácinos puramente serosos. Na glândula labial foi observado ácinos do tipo mistos ou seromucosos. Para a glândula zigomática não foi possível a realização da análise microscópica devido problemas de procedimento histológico. As características das glândulas salivares observadas em G. venusta se assemelham a de outros mamíferos onívoros, entretanto, apresentaram pequenas diferenças no que se refere à sua localização e forma.

The aim of this study was to describe the morphology of the salivary glands of Glironia venusta. A sample of G. venusta was collected in fauna rescue of hidroelectric usine of Teles Pires was used. This exemplar was fixed with formaldehyde solution 10%. For macroscopic analysis, the glands were dissected and photographed in situ and for the microscopy fragments of the glands were removed, that were dehydrated in increasing alcohol concentrations, diaphanizated in xylene, included in paraffin and stained with hematoxylin and eosin. G. venusta presented the mandibular, parotid, sublingual, labial and zygomatic glands. The mandible is shown as elongated structures, composed of non-septated lobes and located in the anterior cervical region. With a triangular format, the parotid was located in the depression of the masseter, ventral an auricular cartilage. The sublingual were located ventral to the mandible, caudal to the digastric muscle. In the posterior dorsal portion, until the labial commissure, dorsal to the orbicularis muscle of mouth was observed the labial glands, which showed a "U" shape. The zygomatic glands was accompanied the shape of the anterior portion of the zygomatic arch at the infraorbital margin. The mandibulars and sublinguals glands showed acini of mucous type, with some serous acini. The parotid one was made up purely by serous acini. In the labial gland was observed acini of mixed types mixed or seromucosous. Related the zygomatic gland was not possible to perform the microscopic analysis because of histological proceeding problems. Characteristics of the salivary glands observed in G. venusta resemble other omnivores mammals, but showed small differences as regards their location and shape.

Caracterização comparativa do intestino das espécies da Ordem Xenarthra / Comparative characterization of the intestine of species from the Xenarthra Order

O sucesso na manutenção de uma espécie depende de vários fatores entre eles a eficiência digestiva, sendo assim parâmetros morfométricos do tubo digestório são necessários para o conhecimento dos processos digestivos dos alimentos no organismo animal além de indicar a preferência alimentar de uma espécie. Este trabalho visou descrever morfologicamente os intestinos delgado e grosso, órgãos do sistema digestório de representantes da ordem Xenarthra a fim de fornecer subsídios para a avaliação da dieta e realização de procedimentos clínicos nestes animais, sejam eles de vida livre ou de cativeiro. Foram utilizados 7 espécimes entre preguiças-de-coleira (Bradypus torquatus), tatu-verdadeiro (Dasypus novemcinctus) e tamanduá-bandeira (Myrmecophaga tridactyla). Todos as amostras foram processadas seguindo procedimentos de rotina efetuados nos laboratórios de Anatomia Animal e Histologia da FZEA/USP. Os intestinos de B. torquatus se apresentaram curtos e simples, enquanto que nos exemplares de D. novemcintus e M. tridactyla o intestino era longo e com algumas peculiaridades. No duodeno de todos os espécimes notamos a presença das glândulas de Brünner e estruturas para aumentar a superfície de absorção. Apenas em preguiças, o mesentério mantém o jejuno preso à parede dorsal da cavidade abdominal. O íleo representou a menor porção nas preguiças e tatus, exceto em tamanduáque apresentava o íleo como a maior parte depois do jejuno. O ceco em tatus e tamanduás apresentavam tamanho considerável e a presença de glândulas na mucosa, nestas espécies destacamos a funcionalidade do ceco, uma vez que este se apresentou repleto de restos alimentares. Na mucosa do cólon de todos os espécimes, haviam criptas de Lieberkühn, sendo mais numerosas em D. novemcinctus e M. tridactyla. Apenas em B. torquatus, o reto apresentou maior diâmetro e rigidez em relação ao cólon. No reto de todas as espécies estudadas, a superfície glandular era numerosa e com grande quantidade de células caliciformes, que produzem muco, para facilitar a defecação. Nossos resultados demonstram que a conformação e estruturas do sistema digestório reflete mais o tipo de alimentação e necessidades digestórias do animal do que à família que ele pertence.(AU)

Morphometric parameters of the digestive tract are required for an understanding of the digestive processes of the food in the animal organism, besides indicating the feeding preference of specie. This study aimed to describe morphologically the small and large intestines, organs of the digestive system of representatives of Xenarthra order to provide data for the evaluation of diet and conduct clinical procedures in these animals, whether free-living or captive. At this research, were used in total 7 specimens from three-toed sloths (Bradypus torquatus), nine-banded armadillo (Dasypus novemcinctus) and giant anteater (Myrmecophaga tridactyla). The intestines of B. torquatus were short and simple, but at the specimens of D. novemcintus and M. tridactyla the intestines were long and had some peculiarities. We notice the presence of Brunner's glands and structures to increase the surface absorption at the duodenum of all specimens. Only in B. torquatus, we notice that the mesentery remains the jejune attached to the dorsal wall of the abdominal cavity. The ileum represented the lower portion of the intestines in all studied specimens except in M.tridactyla. The cecum in D. novemcinctus and M. tridactyla showed considerable size, glands at the mucosa and was full of food debris, indicating that it was functional. In the mucosa of the colon of all specimens had crypts of Lieberkühn, being more numerous in D. novemcinctus and M. tridactyla. Only in B. torquatus, the rectum showed greater diameter and stiffness compared to the colon. In all species studied, we notice a large glandular surface and lots of goblet cells that produce mucus to facilitate defecation. Our results demonstrate that the conformation and structure of the digestive system reflects more the kind of diet and digestive needs of the animal, than to the family he belongs.(AU)

Kinematic gait analyses in healthy Golden Retrievers / Análise cinemática da marcha de cães Golden Retriever saudáveis

Kinematic analysis relates to the relative movement between rigid bodies and finds application in gait analysis and other body movements, interpretation of their data when there is change, determines the choice of treatment to be instituted. The objective of this study was to standardize the march of Dog Golden Retriever Healthy to assist in the diagnosis and treatment of musculoskeletal disorders. We used a kinematic analysis system to analyse the gait of seven dogs Golden Retriever, female, aged between 2 and 4 years, weighing 21.5 to 28 kg, clinically normal. Flexion and extension were described for shoulder, elbow, carpal, hip, femorotibialis and tarsal joints. The gait was characterized lateral and had accepted hypothesis of normality for all variables, except for the stance of hip and elbow, considering a confidence level of 95%, significance level α = 0.05. Variations have been attributed to displacement of the stripes during movement and the duplicated number of reviews. The kinematic analysis proved to be a consistent method of evaluation of the movement during canine gait and the data can be used in the diagnosis and evaluation of canine gait in comparison to other studies and treatment of dogs with musculoskeletal disorders.

A análise cinemática relaciona-se com o movimento relativo entre corpos rígidos e encontra aplicação na análise da marcha e de outros movimentos do corpo. A interpretação de seus dados, quando há alteração, determina a escolha do tratamento a ser instituído. O objetivo deste estudo foi padronizar a marcha do cão Golden Retriever saudável visando auxiliar no diagnóstico e tratamento de afecções músculo esquelética. Neste estudo utilizou-se um sistema de análise cinemática para analisar a marcha de sete cães da raça Golden Retriever, fêmeas, idade entre 2 e 4 anos, peso variando de 21.5 a 28 kg, clinicamente sadias. Dados morfométricos foram coletados para descrever a população estudada. Variáveis de tempo e distâncias foram mensuradas para descrever a marcha, movimentos de flexão e extensão foram descritos para as articulações do ombro, cubital, cárpica, do quadril, femorotibial e társica. A marcha foi caracterizada lateral e teve hipótese de normalidade aceita para todas as variáveis, exceto para o apoio de quadril e apoio de cúbito, considerando um grau de confiança de 95%, ou seja, nível de significância α = 0.05. As variações foram atribuídas ao deslocamento das tarjas durante o movimento e ao repetido número de avaliações. A análise cinemática provou ser um consistente método de avaliação do movimento durante a marcha canina e os dados obtidos podem ser utilizados no diagnóstico e na comparação em avaliações de marcha para outros estudos e tratamento de cães com afecções musculoesqueléticas.

Cat amniotic membrane multipotent cells are nontumorigenic and are safe for use in cell transplantation.

Amnion-derived mesenchymal stem cells (AMSCs) are multipotent cells with an enhanced ability to differentiate into multiple lineages. AMSCs can be acquired through noninvasive methods, and therefore are exempt from the typical ethical issues surrounding stem cell use. The objective of this study was to isolate and characterize AMSCs from a cat amniotic membrane for future application in regenerative medicine. The cat AMSCs were harvested after mechanical and enzymatic digestion of amnion. In culture medium, the cat AMSCs adhered to a plastic culture dish and displayed a fibroblast-like morphology. Immunophenotyping assays were positive for the mesenchymal stem cell-specific markers CD73 and CD90 but not the hematopoietic markers CD34, CD45, and CD79. Under appropriate conditions, the cat AMSCs differentiated into osteogenic, chondrogenic, and adipogenic cell lineages. One advantage of cat AMSCs was nonteratogenicity, assessed 4 weeks post injection of undifferentiated AMSCs into immunodeficient mice. These findings suggest that cat amniotic membranes may be an important and useful source of mesenchymal stem cells for clinical applications, especially for cell or tissue replacement in chronic and degenerative diseases.

Alterações morfofuncionais renais em cães Golden Retriever Distróficos (GRMD) / Kidneys morphological changes in Golden Retriever Muscular Dystrophy dogs (GRMD)

A Golden Retriever Muscular Dystrophy (GRMD) é geneticamente homóloga à distrofia muscular de Duchenne (DMD) que acomete seres humanos. É uma doença genética que gera degeneração progressiva da musculatura esquelética. Considerando-se as intensas alterações musculares, é natural pensar em uma possível lesão renal decorrente da intensa lesão muscular. Foram avaliados seis cães machos da raça Golden Retriever afetados pela distrofia muscular (GRMD) e três cães machos clinicamente sadios. A concentração de creatinina foi determinada e as proteínas urinárias foram avaliadas por eletroforese em gel de poliacrilamida. Os resultados mostraram que a proteinúria patológica não está diretamente associada à Distrofia Muscular de Duchenne, porém diversos parâmetros apresentaram concentrações aumentadas para animais afetados, como a razão proteína/albumina, que foi maior em cães distróficos, podendo ser indício de microalbuminúria e conseqüente lesão renal precoce. Estes resultados visam embasar avaliações clínicas e futuros estudos considerando-se as patologias decorrentes ou associadas a esta doença genética.

The Golden Retriever Muscular Dystrophy (GRMD) is genetically homologous to Duchenne muscular dystrophy (DMD) that affects humans. It is a genetic disease that causes progressive degeneration of skeletal muscle. Considering the intense muscle changes, it is natural to think in possible kidney damage caused by intense muscle injury. We evaluated six male Golden Retriever dogs affected by Duchenne muscular dystrophy (GRMD) and three clinically healthy male dogs. The urinary proteins and creatinine concentration were determined. The proteins were analyzed by polyacrylamide gel electrophoresis. The results showed that pathological proteinuria is not directly associated with Duchenne muscular dystrophy, but several parameters showed increased concentrations for affected animals, as the ratio protein/albumin, which was higher in dystrophic dogs, probably a consequence of microalbuminuria a sign of early kidney damage. These results aim to base future studies and clinical evaluations considering the pathologies arising from or associated with this genetic disease.

Identificação do nicho de progenitores mesenquimais no fígado de embriões e fetos caninos: uma fonte de células-tronco para terapia celular / Identification of mesenchymal progenitor niches from liver of embryo and fetuses of canines: a source of stem cells for cell therapy

As células-tronco (CT) derivadas dos tecidos fetais (TF) foram as mais recentes descobertas entre as CT, e ultimamente tem demonstrado amplo potencial terapêutico, dentre os TF o fígado fetal (FF) apresenta grande potencial terapêutico. Este órgão durante o período fetal em mamíferos é um nicho hematopoético transitório, sendo o principal órgão responsável pela hematopoese no feto, além de contribuir com a formação do nicho definitivo na medula óssea adulta, portanto pode ser considerado um nicho de células-tronco mesenquimais (CTM) e progenitores. No entanto, pouco se sabe sobre a localização destas células no FF, desta forma o presente estudo visa identificar o nicho de CTM e progenitores em FF de cães, a fim de contribuir com as técnicas de isolamento e extração celular. Em conjunto foi realizada a verificação da expressão do fator de transcrição Oct-3/4 e da proteína delta polimerase do DNA (PCNA). Para a análise foram utilizados cinco embriões e 11 fetos caninos com idades gestacionais variando de 25-60 dias. Os resultados elucidaram a partir de 25 dias de gestação o FF apresentou-se volumoso e composto por todas as estruturas típicas, dentre elas a tríade portal, ductos biliares e ramos das artérias hepáticas. Com 30 dias de gestação foram identificados os primeiros sitos de progenitores mesenquimais (PM) enquanto que aos 60 dias os nichos estavam completamente formados com localização semelhante ao fígado adulto (FA). No entanto, células imunopositivas para Oct-3/4 não foram identificadas; sendo assim, destacamos que o FF é uma fonte de PM, apresentando-se como uma alternativa para a utilização terapêutica, bem como para os estudos da biologia do desenvolvimento das CTM e progenitores.(AU)

Stem cells (SC) derived from fetal tissues (FT) were the most recent discoveries among the SC, and lately had demonstrated broad therapeutic potential; the FT from the fetal liver (FL) presents great therapeutic potential. This organ during the fetal period in mammals acts as a transient hematopoietic niche, being the main organ responsible for hematopoiesis in the fetus, and contribute to the formation of permanent niche in the adult bone marrow, thus can be considered a niche for mesenchymal stem cells (MSC) and parents. However, little is known about the location of these cells in FF, so the present study aims to identify the niche of mesenchymal progenitors in FF and dogs in order to contribute to the techniques of cell isolation and extraction. Together was performed to verify the expression of the transcription factor Oct-3/4 of the protein and DNA polymerase delta (PCNA). For the analysis of five embryos were used and 11 canine fetuses with gestational ages ranging from 25-60 days. The results elucidated from 25 days of gestation showed the FF is bulky and composed of all the typical structures, among them the portal triad, bile ducts and hepatic artery branches. With 30 days of gestation were identified the first requirements of mesenchymal progenitors (MP) at 60 days while the niches were completely formed with location similar to adult liver (AL). However, cells immunoreactives for Oct-3/4 were not identified, therefore, point out that the FL is a source of PM, presenting as an alternative for therapeutic purposes as well as for studying the developmental biology of MSC and parents.(AU)

Forma e função dos músculos perineais da viscacha (Lagostomus maximus) / Shape and function of the perineal muscles of viscacha (Lagostomus maximus)

Entre as espécies de roedores já estudadas podemos destacar a grande variação na morfologia do aparelho reprodutor masculino. Assim, considerando a importância ecológica dos roedores, e a grande representatividade numérica quanto geográfica destes animais, bem como a escassez que aborda a anatomia reprodutiva, desenvolvemos este trabalho com um roedor histricomorfo da América do Sul, a viscacha. Este animal apresenta algumas características reprodutivas bem peculiares, desta forma descreveremos a anatomia macroscópica da musculatura perineal, e o papel dos músculos no comportamento copulatório desta espécie. A região perineal da viscacha é composta por cinco músculos, sendo que três músculos encontram-se dispostos no diafragama urogenital superficial, Musculus ischiocavernosus, M. bulbocavernosus e M. bulbospongiosus, e pelos músculos que se encontram no diafragma pélvico, M. levator ani e M. retractor penis. Sendo assim, destacamos que o estudo do assoalho pélvico em animais silvestres é de grande valia, uma vez que contribuem com o melhor entendimento dos mecanismos relacionados â ereção e ejaculação; ou seja colaboram com os estudos sobre a reprodução dos animais.

Among the rodent species studied we can highlight the wide variation in the morphology of the male reproductive system. Thus, considering the ecological importance of rodents, and the large number and geographical representation of this animal, as well as shortages regarding the reproductive anatomy, we developed this study with viscacha, a South American histricomorph rodent. As this species has some very peculiar reproductive features, we described the gross anatomy of the perineal muscles and the role of copulatory behavior. The perineal region of viscacha is composed of five muscles, three of which are arranged in the superficial genitourinary diaphragm, as Musculus ischiocavernosus, M. bulbocavernosus and M. bulbospongiosus, and the muscles that lie at the pelvic diaphragm, M. levator ani and M. retractor penis. Therefore, we emphasize that the study of the pelvic floor in wild animals is of great value, then contribute to a better understanding of the mechanisms related to erection and ejaculation or collaborate with studies on the reproduction of animals.

Morphological characterization of the progenitor blood cells in canine and feline umbilical cord.

The umbilical cord blood (UCB) is an important source of hematopoietic stem cells with great deal of interest in regenerative medicine. The UCB cells have been extensively studied as an alternative to the bone marrow transplants. The challenge is to define specific methods to purify and characterize these cells in different animal species. This study is aimed at morphological characterization of progenitor cells derived from UCB highlighting relevant differences with peripheral blood of adult in dog and cats. Therefore, blood was collected from 18 dogs and 5 cats' umbilical cords from fetus in various developmental stages. The mononuclear cells were separated using the gradient of density Histopaque-1077. Characterization of CD34+ cells was performed by flow cytometric analysis and transmission electron microscopy. Granulocytes (ancestry of the basophiles, eosinophiles, and neutrophiles) and agranulocytes (represented by immature lymphocytes) were identified. We showed for the first time the ultrastructural features of cat UCB cells.

Células-tronco derivadas de tecido adiposo: isolamento, cultivo in vitro e perspectivas de utilização em dermatologia / Adipose tissue derived stem cells: isolation, in vitro culture and potential uses in dermatology

É extensa a literatura existente sobre as características e potenciais terapêuticos das células-tronco mesenquimais adultas, e a possibilidade de seu isolamento a partir de tecido adiposo excedente de outros procedimentos é promissora e extremamente desejável. Os protocolos para tal utilização, porém, ainda são descritos de maneira sumarizada e não padronizada. O objetivo dessa comunicação é apresentar um protocolo simples, prático e eficaz tanto da coleta do tecido adiposo em ambiente hospitalar quanto do isolamento e cultivo, em laboratório, in vitro de células-tronco mesenquimais adultas derivadas de tecido adiposo, com a finalidade de discutir as perspectivas da utilização dessa técnica na dermatologia.

The use of animal models for stroke research: a review.

Stroke has been identified as the second leading cause of death worldwide. Stroke is a focal neurologic deficit caused by a change in cerebral circulation. The use of animal models in recent years has improved our understanding of the physiopathology of this disease. Rats and mice are the most commonly used stroke models, but the demand for larger models, such as rabbits and even nonhuman primates, is increasing so as to better understand the disease and its treatment. Although the basic mechanisms of stroke are nearly identical among mammals, we here discuss the differences between the human encephalon and various animals. In addition, we compare common surgical techniques used to induce animal models of stroke. A more complete anatomic knowledge of the cerebral vessels of various model species is needed to develop more reliable models for objective results that improve knowledge of the pathology of stroke in both human and veterinary medicine.